Discussion

Flowcytometry has proven enormous potential for personalized medicine as well as for clinical drug development and project decision making, because in many disease areas the therapeutic options and the drug mode of action can only be understood on a cellular level. Consequently, flowcytometry and is considered an emerging field by the industry (e.g. Ligand Binding Assay Bioanalytical Focus Group of the American Association of Pharmaceutical Scientists meeting, NBC; San Diego, May 2014). Nevertheless, in a clinical setting conventional flowcytometry today often is regarded being cumbersome, costly and potentially error prone due to the very limited biomarker stability on and in cells.
Chipcytometry aims at standardizing the overall workflow by improving storage capabilities of biomarkers while maintaining full cellular integrity. This significantly expands the multiplexing capabilities and provides actionable high-content cytometry data output. The Chipcytometry technology first has been described by Hennig et al. 2008 and is based on iterative imaging cytometry. Chipcytometry allows preserving biomarkers on intact cells and tissues for more than 12 month allowing re-staining samples again and again due to immobilization of the cells within microfluidic chips. The platform has been validated for a range of specimens including whole blood, PBMC, CSF and various tissue types. After immobilization (and optional fixation using fixatives like PFA), a virtually unlimited set of markers can be analyzed or a broad range of functional tests based on cyclic staining/imaging/switch-off of remaining fluorescence. Chipcytometry for the first time enables long-term cell storage combined with 50-plex analysis of immune cells and delivering easy to access high-content cytometry data based on innovative data-visualization tools and compatible to industry standard software like FloJo (Treestar Inc.) and Watson LIMS (Thermo Scientific Inc.).
In this study we present data from three clinical biomarkers panels validated according to industry guidelines. The studies focus on multiple sclerosis biomarkers, immune-oncology biomarkers and autoimmune diseases biomarkers. Finally, time and effort is benchmarked with other cytometry technologies.
Chipcytometry was developed by Hannover Medical School, Department of Pediatric Pneumology, Allergology and Neonatology and under a grant by Go-Bio/ Germany Research Ministry.